LOCUS       sequence-418117-        3611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV HDR donor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3611)
  AUTHORS   Tycko J, Van MV, Aradhana, DelRosso N, Ye H, Yao D, Valbuena R, 
            Vaughan-Jackson A, Xu X, Ludwig C, Spees K, Liu K, Gu M, Khare V, 
            Mukund AX, Suzuki PH, Arana S, Zhang C, Du PP, Ornstein TS, Hess GT,
            Kamber RA, Qi LS, Khalil AS, Bintu L, Bassik MC
  TITLE     Development of compact transcriptional effectors using 
            high-throughput measurements in diverse contexts.
  JOURNAL   Nat Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6.
  PUBMED    39487265
REFERENCE   2  (bases 1 to 3611)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6."
COMMENT     Sequence Label: pJT310 AAV-BFPdonor
FEATURES             Location/Qualifiers
     source          1..3611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      98..553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(185..204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     395..416
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(570..589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     683..705
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(737..755)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        823..927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(928..996,997..1788)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1146..1165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1959..2547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2448..2467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2609..2738
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(2762..2764,2765..3472)
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     CDS             join(2762..2764,2765..3472)
                     /codon_start=1
                     /product="blue variant of GFP"
                     /label=BFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     CDS             join(2762..2764,2765..3472)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(2804..2825)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3065..3084)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3412..3433
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   join(3494..3611,1..23)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(3494..3611,1..12)
                     /label=AAV2 ITR
ORIGIN
        1 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
       61 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
      121 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
      181 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      241 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      301 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      361 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
      421 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
      481 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
      541 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      601 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
      661 tccgcttaca gacaagctgt gaccgggagc tgcatgtgtc agaggttttc accgtcatca
      721 ccgaaacgcg caaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
      781 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      841 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      901 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      961 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1021 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1081 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1141 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1201 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1261 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1321 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1381 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1441 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1501 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1561 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1621 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1681 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1741 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1801 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1861 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1921 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1981 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2041 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2101 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2161 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2221 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2281 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2341 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2401 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2461 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2521 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2581 gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact
     2641 gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     2701 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc gtgtgtctag
     2761 agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2821 cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg
     2881 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2941 cgtgaccacc ctgagccacg gggtgcagtg cttcagccgc taccccgacc acatgaagca
     3001 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3061 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3121 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3181 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     3241 catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     3301 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3361 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3421 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acagtaacgg
     3481 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3541 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3601 tcagtgagcg a
//
